LOCUS       sequence-213210-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Met-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7712)
  AUTHORS   Villarreal F, Contreras-Llano LE, Chavez M, Ding Y, Fan J, Pan T, 
            Tan C
  TITLE     Synthetic microbial consortia enable rapid assembly of pure 
            translation machinery.
  JOURNAL   Nat Chem Biol. 2018 Jan;14(1):29-35. doi: 10.1038/nchembio.2514. 
            Epub 2017 Nov 13.
  PUBMED    29131146
REFERENCE   2  (bases 1 to 7712)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nchembio.2514"; journalName: "Nat Chem Biol"; date: 
            "2018-01"; volume: "14"; issue: "1"; pages: "29-35"
COMMENT     Sequence Label: B1-2
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     171..190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     274..293
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        274..292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             332..354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2395..2412
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2486..2504)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2500..2547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2722..2727,2728..2744,2745..2750))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2777..2795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2863..2967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2968..3036,3037..3828)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3186..3205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3999..4587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4488..4507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4741..4758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4773..4912
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4838..4857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4998..5020
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5014..5205)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6517..7599)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7561..7580
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7600..7677)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
       61 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
      121 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
      181 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      241 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattga
      301 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      361 atgcatactc aagtcgcgaa gaaaattctg gtgacgtgcg cactgccgta cgctaacggc
      421 tcaatccacc tcggccatat gctggagcac atccaggctg atgtctgggt ccgttaccag
      481 cgaatgcgcg gccacgaggt caacttcatc tgcgccgacg atgcccacgg tacaccgatc
      541 atgctgaaag ctcagcagct tggtatcacc ccggagcaga tgattggcga aatgagtcag
      601 gagcatcaga ctgatttcgc aggctttaac atcagctatg acaactatca ctcgacgcac
      661 agcgaagaga accgccagtt gtcagaactt atctactctc gcctgaaaga aaacggtttt
      721 attaaaaacc gcaccatctc tcagctgtac gatccggaaa aaggcatgtt cctgccggac
      781 cgttttgtga aaggcacctg cccgaaatgt aaatccccgg atcaatacgg cgataactgc
      841 gaagtctgcg gcgcgaccta cagcccgact gaactgatcg agccgaaatc ggtggtttct
      901 ggcgctacgc cggtaatgcg tgattctgaa cacttcttct ttgatctgcc ctctttcagc
      961 gaaatgttgc aggcatggac ccgcagcggt gcgttgcagg agcaggtggc aaataaaatg
     1021 caggagtggt ttgaatctgg cctgcaacag tgggatatct cccgcgacgc cccttacttc
     1081 ggttttgaaa ttccgaacgc gccgggcaaa tatttctacg tctggctgga cgcaccgatt
     1141 ggctacatgg gttctttcaa gaatctgtgc gacaagcgcg gcgacagcgt aagcttcgat
     1201 gaatactgga agaaagactc caccgccgag ctgtaccact tcatcggtaa agatattgtt
     1261 tacttccaca gcctgttctg gcctgccatg ctggaaggca gcaacttccg caagccgtcc
     1321 aacctgtttg ttcatggcta tgtgacggtg aacggcgcaa agatgtccaa gtctcgcggc
     1381 acctttatta aagccagcac ctggctgaat cattttgacg cagacagcct gcgttactac
     1441 tacactgcga aactctcttc gcgcattgat gatatcgatc tcaacctgga agatttcgtt
     1501 cagcgtgtga atgccgatat cgttaacaaa gtggttaacc tggcctcccg taatgcgggc
     1561 tttatcaaca agcgttttga cggcgtgctg gcaagcgaac tggctgaccc gcagttgtac
     1621 aaaaccttca ctgatgccgc tgaagtgatt ggtgaagcgt gggaaagccg tgaatttggt
     1681 aaagccgtgc gcgaaatcat ggcgctggct gatctggcta accgctatgt cgatgaacag
     1741 gctccgtggg tggtggcgaa acaggaaggc cgcgatgccg acctgcaggc aatttgctca
     1801 atgggcatca acctgttccg cgtgctgatg acttacctga agccggtact gccgaaactg
     1861 accgagcgtg cagaagcatt cctcaatacg gaactgacct gggatggtat ccagcaaccg
     1921 ctgctgggcc acaaagtgaa tccgttcaag gcgctgtata accgcatcga tatgaggcag
     1981 gttgaagcac tggtggaagc ctctaaagaa gaagtaaaag ccgctgccgc gccggtaact
     2041 ggcccgctgg cagatgatcc gattcaggaa accatcacct ttgacgactt cgctaaagtt
     2101 gacctgcgcg tggcgctgat tgaaaacgca gagtttgttg aaggttctga caaactgctg
     2161 cgcctgacgc tggatctcgg cggtgaaaaa cgcaatgtct tctccggtat tcgttctgct
     2221 tacccggatc cgcaggcact gattggtcgt cacaccatta tggtggctaa cctggcacca
     2281 cgtaaaatgc gcttcggtat ctctgaaggc atggtgatgg ctgccggtcc tggcgggaaa
     2341 gatattttcc tgctaagccc ggatgccggt gctaaaccgg gtcatcaggt gaaacatcat
     2401 catcaccatc attaaagcag aggccatatg ctcgaggatc cggctgctaa caaagcccga
     2461 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2521 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca
     2581 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     2641 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     2701 ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat
     2761 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     2821 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2941 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3181 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     3241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3481 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     3541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4141 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4681 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4741 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4801 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     4861 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4921 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4981 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5041 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5101 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     5161 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     5221 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     5281 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     5341 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     5401 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     5461 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     5521 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     5581 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     5641 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     5701 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     5761 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     5821 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     5881 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     5941 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     6001 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     6061 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     6121 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     6181 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     6241 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     6301 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     6361 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     6421 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     6481 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6601 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     6661 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     6721 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     6781 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6841 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     6901 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     6961 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     7021 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     7081 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     7141 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     7201 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     7261 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     7321 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     7381 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     7441 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     7501 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     7561 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     7621 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     7681 gatctcgacg ctctccctta tgcgactcct gc
//
